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Double Strandedness of Ribonucleic Acid of Bovine Ephemeral Fever Virus
Author(s) -
Tanaka Yoshio,
Inaba Yuji,
Ito Yasuichiro,
Sato Kunihiko,
Omori Tsuneyoshi,
Matumoto Minoru
Publication year - 1972
Publication title -
japanese journal of microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0021-5139
DOI - 10.1111/j.1348-0421.1972.tb00634.x
Subject(s) - virus , infectivity , rna , biology , centrifugation , rnase p , virology , negative stain , staining , differential centrifugation , microbiology and biotechnology , nucleic acid , chemistry , biochemistry , electron microscope , gene , physics , genetics , optics
Virus labeled with 3 H‐uridine or 32 P‐orthophosphate was purified by CsCl equilibrium centrifugation of concentrated virus materials from infectious BHK21‐WI2 cell culture fluids. A single clearly visible band formed in the gradient coinciding with a sharp peak of radioactivity having a buoyant density of 1.19 g/ml. Infectivity exhibited a broader distribution with a peak coinciding with the visible band, in which numerous virions of the virus were observed with the aid of an electron microscope using the phosphotungstic negative staining technique. Ribonucleic acid (RNA) extracted from the purified virus by the phenol method exhibited a rather broad distribution of radioactivity with a major peak at about 12 S when analyzed by the sucrose density gradient centrifugation technique. Viral RNA centrifuged in the gradient after ribonuclcase (RNase) treatment showed a single sharp peak at about 12 S. These findings seemed to indicate that the virion of this virus contained double‐stranded RNA. Double strandedness of the viral RNA was further corroborated by an examination of the base composition, reduced resistance to RNase in low salt concentration and a sharp thermal transition with a relatively high melting temperature of 85 C. The virus could not be classified either in the rhabdovirus group, although the shape of the virion resembled that of the rhabdoviruses, or in the reovirus group since the virus was ether‐sensitive. It seemed necessary to create a new genus for this virus.